CUSTOM REAL-TIME/QUANTITATIVE
TaqmanTM PCR (QPCR Form D-2)
Please print and complete this form and forward/fax to MDS. We will contact you for a detailed quotation.
Institution:
Investigator:
Email:
Billing Address:
Submitted by:
Date Submitted:
/
/
PO #:
Shipping Address:
Zip:
Fax:(
State:
)
-
Phone:(
Services Requested
Complete PCR analysis (continue)
City:
)
-
Ext::
Plate reading only (complete Form D-4)
Please provide us with an overview of your project:
Target
1. Are you interested in quantifying a DNA target or RNA target (requires an RT reaction)
2. What is the target?
3. Will you provide the primers and probes? Yes
No
If no, then please fill out the assay development section.
Reagent information (If no assay development is required)
Primer Set #1: Forward
; Concentration to use:
Reverse
; Concentration to use:
Probe Name:
; Concentration to use:
Fluorescent dye:
6-FAM
VIC
TET
Other:
;
Quencher dye:
TAMRA
Other:
Forward
; Concentration to use:
Reverse
; Concentration to use:
Probe Name:
; Concentration to use:
Fluorescent dye:
6-FAM
VIC
TET
Other:
Quencher dye:
TAMRA
Other:
SYBR green
Primer Set #2:
;
SYBR green
Standard information (If no assay development is required)
4. What type of standard will be used?
Absolute
Relative
5. What titration range of the standard would you like performed:
6. How many replicates do you require? Duplicate
Triplicate
to
.
Other
Thermocycle program parameters (If no assay development is required)
7. Pre-Amplification cycles:
Temp
; Time
; Number of cycles
Temp
; Time
; Number of cycles
Temp
; Time
; Number of cycles
8. Amplification Cycle:
Denaturing temp
Annealing temp
Extension temp
Number of cycles
; Time
; Time
; Time
9. Post-amplification cycles:
Temp
; Time
; Number of cycles
Temp
; Time
; Number of cycles
Temp
; Time
; Number of cycles
Assay Development
10. Do you require assay development?
Yes
No
If no, then please be sure to fill out the Reagent information, Standard Information and and
thermocycling parameters sections.
11. How will you provide the sequence of your target?
Disk
Email
Genbank#
12. What is the relative expression level of your target
High copy
Low copy
Unknown
13. If an absolute standard is required, do you have a clone containing the target sequence that
can be used as a DNA/RNA standard
Yes
No
Please provide the size of the plasmid to be used as a standard. DNA
RNA transcription unit
length
14. Are you interested in having the absolute standard tested in a background of nucleic acid to
control for matrix effects?
Yes
No
15. If a relative standard is required (ie normalization to an endogenous control or a designated
sample/calibrator), would you like the calibrator diluted to produce a standard curve or
used as is?
Will you provide the appropriate nucleic acid?
Yes
No
16. Do you need to normalize to a particular control gene Yes
No
17. How many replicates do you require?
Duplicate
Triplicate
Other
Samples (For multiple samples, please complete Form D-3.)
18. Number of samples to be submitted
.
19. What type of sample will be submitted? (see recommendations page 3)
Sample source:
human
rat
mouse
other
Cell culture (flask size
; number of flasks
; passage #
)
Cell pellet (number of cells
; lot #
)
Tissue (type
; amount
)
DNA (concentration
; volume
; total amount
ng)
RNA (concentration
; volume
; total amount
ng)
Lysis buffer (if applicable)
Special components of buffer (if applicable)
20. Do we need to extract the entire sample or can we extract a part of the sample?
Part
Whole
21. What method would you prefer for DNA/RNA isolation (Unless otherwise noted, we will use
Qiagen DNeasy procedure for DNA and Qiagen RNeasy procedure for RNA extraction)
DNA:
Qiagen DNeasy
AmbionTrizol
Other:
RNA:
Qiagen RNeasy
AmbionTrizol
Other:
22. Is the sample derived from the primate cells or tissue (human xenograft on mouse, etc.)?
Yes
No (This is an important safety issue.)
Shipping Conditions
23. How do you plan on shipping the samples?
Ambient temperature
Wet/blue ice
Dry ice
Recommend Sample Size for Endpoint PCR
Sample
Amount*
Cultured cells
5 x 10^6
Liver
25 mg (~2mm3)
Brain+
25 mg
Lung+
25 mg
Heart/muscle+
25 mg
Kidney
25 mg
Spleen+
10 mg
Plant leaf
100 mg
Mouse tail
0.8-1.2 cm
Blood
1 ml
Buffy coat
50 ul
DNA
50 ng/reaction
RNA
100 ng RNA/reaction
Other
Call
+These are difficult tissues and may require special processing.
Recommended Shipping Conditions
Plasmid DNA
Liquid = A, W, D; pellet A
PCR products
Liquid = A, W, D; pellet A
Primers
Liquid = A, W, D; pellet A
cDNA or genomic libraries
With DMSO = D; without DMSO = W
RNA
D
Bacterial colonies
A, W
Stab culture
A
Cell banks
D
Glycerol stocks
D
Tissues
D
Blood
W
A = ambient temperature W = wet ice D = dry ice